WO2023224418A1 - Method for producing from fatty alcohols monomers for producing various synthetic resins - Google Patents

Abstract

The present invention relates to a method for producing, from fatty alcohols, various monomers that are used in the production of synthetic resins.

Description

Method for producing various monomers for manufacturing synthetic resins from fatty alcohols

The present invention relates to a method of producing various monomers used in the production of synthetic resins from fatty alcohols.

Bioplatform compounds are produced through biological or chemical conversion based on biomass-derived raw materials and are used for the synthesis of polymer monomers and new materials.

Among bioplatform compounds, hydroxyalkanoic acid, aminoalkanoic acid, alkanediol, aminoalkanol, diaminoalkane, etc. are substances used as monomers for producing synthetic resins such as polyamide and polyester. However, in the case of alkane or alkanol compounds, which are raw materials for manufacturing these bioplatform compounds, if they are short chains with 6 or less carbon atoms, they cause cytotoxicity. Therefore, using these short chain alkane compounds, the above-mentioned biological methods can be used. There was a problem in that it was actually difficult to manufacture bioplatform compounds.

Therefore, there is a need for research or development of technologies that can biologically produce short-chain bioplatform compounds.

The purpose of the present invention is to provide a method for producing monomers for producing synthetic resins from fatty alcohols.

In order to achieve the above object, one aspect of the present invention includes the steps of synthesizing a dialkyl ether compound from fatty alcohol; Fermenting the dialkyl ether compound with a genetically recombinant transformant to produce a dialkyl ether compound functionalized at both ends; and decomposing the dialkyl ether compound functionalized at both ends of the fatty alcohol.

In addition, in order to achieve the above object, another aspect of the present invention includes synthesizing an asymmetric fatty ether compound from fatty alcohol; fermenting the asymmetric fatty ether compound with a genetically recombinant transformant to produce an asymmetric fatty ether compound functionalized at both ends; and decomposing the asymmetric fatty ether compound functionalized at both ends. It provides a method for producing a monomer for producing a synthetic resin from fatty alcohol, including a step.

When a fatty alcohol is transformed into a dialkyl ether compound and used as a substrate, as in the present invention, various types of monomers in which both ends are carboxylated, hydroxylated, or aminated are obtained through a process of hydrolyzing the functionalized product. Two equivalents can be produced, and functionalization is possible even in the case of short-chain fatty alcohols, which are previously difficult to functionalize, and can be manufactured with different functional groups at both ends. Therefore, there is an advantage in that monomers for producing various types of synthetic resins can be easily produced compared to the prior art.

However, the effects of the present invention are not limited to the effects mentioned above, and other effects not mentioned will be clearly understood by those skilled in the art from the following description.

Figure 1 schematically illustrates the entire process of the present invention for producing monomers for producing synthetic resins from fatty alcohols.

Figure 2a is a graph showing the results of synthesizing dioctyl ether with bio-derived 1-octanol, and Figure 2b is a graph showing the results confirming the cytotoxicity of dioctyl ether produced as above.

Figure 2c is a graph showing the results of synthesizing didodecyl ether with bio-derived 1-dodecanol, and Figure 2d is a graph showing the results confirming the cytotoxicity of didodecyl ether produced as above.

Figure 3 is a graph showing the results of confirming the cytotoxicity of dialkyl ether compounds with various carbon numbers against the α,ψ-carboxylated recombinant strain of the present invention.

Figures 4a and 4b show that dioctyl ether produced from bio-derived 1-octanol and didodecyl ether synthesized from bio-derived 1-dodecanol, respectively, by the α,ψ-carboxylated recombinant strain of the present invention. This graph confirms the result of both terminals being functionalized with carboxyl groups.

Figure 5 is a graph confirming the results of dioctyl ether functionalized with carboxyl groups at both ends functionalized with hydroxy groups by the α,ψ-hydroxylated recombinant strain of the present invention.

Figure 6 is a graph confirming the results of dioctyl ether functionalized with hydroxy groups at both ends being functionalized with amine groups at both ends by the α,ψ-aminated recombinant strain of the present invention.

Figure 7 is a graph showing the results of hydrolysis of dioctyl ether functionalized with carboxyl groups at both ends to produce 2 equivalents of 8-hydroxyoctanoic acid.

Figure 8 is a graph showing the results of hydrolysis of dioctyl ether functionalized with hydroxy groups at both ends to produce 2 equivalents of 1,8-octanediol.

Figure 9 is a graph showing the results of producing carboxymethyl 11-carboxyundecyl ether (=12-(carboxymethoxy)dodecanoic acid), a compound in which both ends of ethyldodecyl ether are carboxylated.

According to one embodiment of the present invention, a first step of synthesizing a dialkyl ether compound from fatty alcohol; A second step of fermenting the dialkyl ether compound with a genetically recombinant transformant to produce a dialkyl ether compound functionalized at both ends; and a third step of decomposing the dialkyl ether compound functionalized at both ends. It relates to a method of producing a monomer for producing a synthetic resin from fatty alcohol, comprising a.

In the first step, the fatty alcohol is a compound in which one end of a saturated hydrocarbon with n carbon atoms is substituted with a hydroxy group. The carbon number n of the hydrocarbon may be an integer of 3 to 20, for example, an integer of 4 to 18, specifically an integer of 5 to 16, especially an integer of 6 to 14. Additionally, the hydrocarbon may be straight chain or branched, but may especially be straight chain.

In the second step, the first genetically recombined transformant is used to oxidize both ends of the dialkyl ether compound produced in the first step to introduce a carboxyl group at the α, ψ-position of the dialkyl ether compound. You can. At this time, the first transformant may be a recombinant transformant capable of ψ-oxidation, for example, the β-oxidation pathway is blocked, CYP450, CPRb (CYP450 reductase complex), FADH (fatty alcohol dehydrogenase), FAO (fatty alcohol dehydrogenase) It may be a microorganism that has been genetically engineered to overexpress alcohol oxidase (FALDH), fatty aldehyde dehydrogenase (FALDH), or a combination thereof.

In the second step, a dialkyl ether compound functionalized with a carboxyl group at both ends is converted into a dialkyl ether compound functionalized with a hydroxy group at both ends using a genetically recombinant second transformant or an appropriate catalyst. You can. The second transformant may be a recombinant transformant capable of ψ-reduction, for example, genetically engineered to overexpress CAR (carboxylic acid reductase), Sfp (4'-phosphopantetheinyl transferase), or a combination thereof. It could be a microorganism.

In the second step, a dialkyl ether compound functionalized with hydroxy groups at both ends is converted into a dialkyl ether compound functionalized with amine groups at both ends using a third genetically recombinant transformant or an appropriate catalyst. You can. The third transformant may be a recombinant transformant capable of ψ-amination, for example, overexpressing ψ-TA (ψ-transaminase), AlaDH (alanine dehydrogenase), ADH (Alcohol dehydrogenase), or a combination thereof. Preferably, it can be a genetically engineered microorganism.

In the third step, various types of monomers such as hydroxyalkanoic acid, aminoalkanoic acid, alkanediol, aminoalkanol, diaminoalkane, etc. can be produced through hydrolysis of the ether bond.

Furthermore, in the present invention, synthetic resins such as polyamide and polyester can be manufactured using various types of monomers produced in the third step. In particular, lactone compounds or lactam compounds can be produced using hydroxyalkanoic acid monomers. can be manufactured.

According to another embodiment of the present invention, a first' step of synthesizing an asymmetric fatty ether compound from fatty alcohol; A second' step of fermenting the asymmetric fatty ether compound with a genetically recombinant transformant to produce an asymmetric fatty ether compound functionalized at both ends; and a third' step of decomposing the asymmetric fatty ether compound functionalized at both ends.

In the first' step, the fatty alcohol is a compound in which one end of a saturated or unsaturated aliphatic hydrocarbon with m or n carbon atoms is substituted with a hydroxy group. The carbon number m of the hydrocarbon may be an integer of 1 to 20, for example, an integer of 1 to 12, specifically an integer of 1 to 6, especially an integer of 1 to 3. In addition, the carbon number n of the hydrocarbon may be an integer of 3 to 20, for example, an integer of 4 to 18, specifically an integer of 5 to 16, especially an integer of 6 to 14. Additionally, the hydrocarbons may be straight chain or branched, but may especially be straight chain.

In the second' step, both ends of the dialkyl ether compound produced in the first step are oxidized using the first genetically recombined transformant to form a carboxyl group at the α, ψ-position of the dialkyl ether compound. can be introduced. At this time, the first transformant may be a recombinant transformant capable of ψ-oxidation, for example, the β-oxidation pathway is blocked, CYP450, CPRb (CYP450 reductase complex), FADH (fatty alcohol dehydrogenase), FAO (fatty alcohol dehydrogenase) It may be a microorganism that has been genetically engineered to overexpress alcohol oxidase (FALDH), fatty aldehyde dehydrogenase (FALDH), or a combination thereof.

In the second' step, a dialkyl ether compound functionalized with a carboxyl group at both ends is converted into a dialkyl ether compound functionalized with a hydroxy group at both ends using a genetically recombinant second transformant or an appropriate catalyst. You can do it. The second transformant may be a recombinant transformant capable of ψ-reduction, for example, genetically engineered to overexpress CAR (carboxylic acid reductase), Sfp (4'-phosphopantetheinyl transferase), or a combination thereof. It could be a microorganism.

In the second' step, a dialkyl ether compound functionalized with hydroxy groups at both ends is converted into a dialkyl ether compound functionalized with amine groups at both ends using a third genetically recombinant transformant or an appropriate catalyst. You can do it. The third transformant may be a recombinant transformant capable of ψ-amination, for example, overexpressing ψ-TA (ψ-transaminase), AlaDH (alanine dehydrogenase), ADH (Alcohol dehydrogenase), or a combination thereof. Preferably, it can be a genetically engineered microorganism.

In the 3' step, various types of monomers such as hydroxyalkanoic acid, aminoalkanoic acid, alkanediol, aminoalkanol, diaminoalkane, etc. can be produced through hydrolysis of the ether bond.

Furthermore, in the present invention, synthetic resins such as polyamide and polyester can be manufactured using various types of monomers produced in the third step. In particular, lactone compounds or lactam compounds can be produced using hydroxyalkanoic acid monomers. can be manufactured.

Hereinafter, the present invention will be described in detail.

One. Embodiment 1 - Strategy for Mediating Symmetric Dialkyl Ether Compounds

One aspect of the present invention provides a method for producing monomers for producing synthetic resins from fatty alcohols.

The method of the present invention includes synthesizing a dialkyl ether compound from fatty alcohol; Fermenting the dialkyl ether compound with a genetically recombinant transformant to produce a dialkyl ether compound functionalized at both ends; and decomposing the dialkyl ether compound functionalized at both ends.

first stage

First, a dialkyl ether compound is synthesized from fatty alcohol.

The fatty alcohol is a compound in which one end of a saturated hydrocarbon with n carbon atoms is substituted with a hydroxy group. The carbon number n of the hydrocarbon may be an integer of 3 to 20, for example, an integer of 4 to 18, specifically an integer of 5 to 16, especially an integer of 6 to 14. Additionally, the hydrocarbon may be straight chain or branched, but may especially be straight chain.

The fatty alcohol described above can be converted into a dialkyl ether compound through a reaction shown in Chemical Formula 1 below. A dialkyl ether compound having 2n carbon atoms can be produced from a fatty alcohol containing n carbon atoms through the process shown below. The dialkyl ether compound produced at this time has the same type of hydrocarbon group with n carbon atoms on both sides of the O atom of the ether bond, so it is called a symmetrical dialkyl ether compound.

[Formula 1]

Therefore, from fatty alcohols such as butanol, pentanol, hexanol, heptanol, octanol, nonanol, decanol, undecanol, dodecanol, etc., dibutyl ether (C ₈ H ₁₈ O) and dipentyl ether (C ₁₀ H ₂₂ O), dihexyl ether (C ₁₂ H ₂₆ O), diheptyl ether (C ₁₄ H ₃₀ O), dioctyl ether (C ₁₆ H ₃₄ O), dinonyl ether (C ₁₈ H ₃₈ O), dide Dialkyl ether compounds such as syl ether (C ₂₀ H ₄₂ O), diundecyl ether (C ₂₂ H ₄₆ O), and didodecyl ether (C ₂₄ H ₅₀ O) can be produced.

In addition, Formula 1 as described above can be carried out through methods well known in the technical field to which the present invention pertains, for example, nucleophilic substitution at a temperature of 130 to 140 degrees Celsius using an acid catalyst such as sulfuric acid (H ₂ SO ₄ ). It can be performed by proceeding with the reaction, but is not limited to this.

second stage

The dialkyl ether compound produced in the first step is fermented with a genetically recombinant transformant to produce a dialkyl ether compound functionalized at both ends.

First, using the first genetically recombined transformant, both ends of the dialkyl ether compound produced in the first step are oxidized to form a carboxyl group at the α, ψ-position of the dialkyl ether compound, as shown in Chemical Formula 2 below. introduce.

[Formula 2]

Therefore, the first transformant may be a recombinant transformant capable of ψ-oxidation, for example, the β-oxidation pathway is blocked, CYP450, CPRb (CYP450 reductase complex), FADH (fatty alcohol dehydrogenase), FAO (fatty alcohol dehydrogenase) It may be a microorganism genetically engineered to overexpress alcohol oxidase (FALDH), fatty aldehyde dehydrogenase (FALDH), or a combination thereof, but is not limited thereto, and a person skilled in the art may use an appropriate microorganism for ψ-oxidation. As long as it is genetically recombined as much as possible, it can be used as the first transformant of the present invention without limitation. Here, the microorganisms include Escherichia, Corynebacterium, Clostridium, Zymonomas, Salmonella, Rhodococcus, Pseudomonas, Bacillus, Lactobacillus, Enterococcus, Alcaligenes, Klesiella, Paenibacillus, Arthrobacter, Brevibacterium , Pichia, Candida, Hansenula, Synechococcus, Synechocystis, Anabaena, Ralstonia, Lactococcus ( It may be a microorganism of the genus Lactococcus or Saccharomyces, but is not limited thereto.

When the dialkyl ether compound is supplied as a substrate to the first transformant for fermentation, the carbon at the ψ-position of the dialkyl ether compound can be oxidized and functionalized into a carboxyl group.

Meanwhile, using a genetically recombinant second transformant or an appropriate catalyst, the dialkyl ether compound produced as above, both ends of which are functionalized with a carboxyl group, is further converted into a dialkyl ether compound, both ends of which are functionalized with a hydroxy group, as shown in the following formula (3). It can be converted to alkyl ether compounds.

[Formula 3]

Therefore, the second transformant may be a recombinant transformant capable of ψ-reduction, and is genetically engineered to overexpress, for example, CAR (carboxylic acid reductase), Sfp (4'-phosphopantetheinyl transferase), or a combination thereof. It may be a microorganism, but is not limited thereto, and if a person skilled in the art of the present invention genetically recombines an appropriate microorganism to enable ψ-reduction, it can be used as the second transformant of the present invention without limitation. . Here, the microorganisms include Escherichia, Corynebacterium, Clostridium, Zymonomas, Salmonella, Rhodococcus, Pseudomonas, Bacillus, Lactobacillus, Enterococcus, Alcaligenes, Klesiella, Paenibacillus, Arthrobacter, Brevibacterium , Pichia, Candida, Hansenula, Synechococcus, Synechocystis, Anabaena, Ralstonia, Lactococcus ( It may be a microorganism of the genus Lactococcus or Saccharomyces, but is not limited thereto.

When the dialkyl ether compound functionalized with carboxyl groups at both ends is supplied as a substrate to the second transformant for fermentation, the carboxyl group at the ψ-position of the dialkyl ether compound functionalized with carboxyl groups at both ends is reduced to a hydroxy group. It can be functionalized.

Furthermore, using a third genetically recombinant transformant or an appropriate catalyst, the dialkyl ether compound produced as above, both ends of which are functionalized with hydroxy groups, can be converted into a dialkyl ether compound, both ends of which are functionalized with amine groups, as shown in the following formula (4). It can be converted to alkyl ether compounds.

[Formula 4]

Therefore, the third transformant may be a recombinant transformant capable of ψ-amination, for example, ψ-TA (ψ-transaminase), AlaDH (alanine dehydrogenase), ADH (Alcohol dehydrogenase), or a combination thereof. It may be a microorganism that has been genetically engineered to overexpress, but is not limited to this, and if a person skilled in the art to which the present invention pertains has genetically recombined an appropriate microorganism to enable ψ-amination, the third transformation of the present invention may be performed without limitation. It can be used as a sieve. Here, the microorganisms include Escherichia, Corynebacterium, Clostridium, Zymonomas, Salmonella, Rhodococcus, Pseudomonas, Bacillus, Lactobacillus, Enterococcus, Alcaligenes, Klesiella, Paenibacillus, Arthrobacter, Brevibacterium , Pichia, Candida, Hansenula, Synechococcus, Synechocystis, Anabaena, Ralstonia, Lactococcus ( It may be a microorganism of the genus Lactococcus or Saccharomyces, but is not limited thereto.

When the dialkyl ether compound functionalized with hydroxy groups at both ends is supplied to the third transformant as a substrate for fermentation, the hydroxy group at the ψ-position of the dialkyl ether compound functionalized with hydroxy groups at both ends is aminated to form an amine. It can be functionalized.

3rd stage

The dialkyl ether compound functionalized at both ends produced in the second step is decomposed. The decomposition is to hydrolyze the ether bond of the dialkyl ether compound functionalized at both ends, and the hydrolysis of the ether bond can be performed through a method well known in the art to which the present invention pertains, for example, sulfuric acid. It can be performed by reacting at a temperature of 200 degrees Celsius using an acid catalyst such as (H ₂ SO ₄ ), but is not limited to this.

Hydrolysis of the ether bond as described above can be performed on all types of dialkyl ether compounds functionalized at both ends that can be produced in step 2. Therefore, a dialkyl ether compound with 2n carbon atoms functionalized with carboxyl groups at both ends is hydrolyzed to produce 2 equivalents of hydroxyalkanoic acid with n carbon atoms or 2 equivalents of aminoalkanoic acid with n carbon atoms, or a dialkyl ether compound with 2n carbon atoms functionalized with a hydroxyl group at both ends is produced. A dialkyl ether compound having 2n carbon atoms is hydrolyzed to produce 2 equivalents of an alkanediol with n carbon atoms or 2 equivalents of an aminoalkanol with n carbon atoms, or a dialkyl ether compound with 2n carbon atoms functionalized with amine groups at both ends is hydrolyzed. By decomposition, 2 equivalents of aminoalkanol with n carbon atoms or 2 equivalents of diaminoalkane with n carbon atoms can be produced.

Various types of monomers such as hydroxyalkanoic acid, aminoalkanoic acid, alkanediol, aminoalkanol, diaminoalkane, etc. prepared as described above can be usefully used to produce synthetic resins such as polyamide and polyester. In particular, hydroxyalkanoic acids are lactone compounds through reactions known in previously known literature (e.g., Pyo et al . (2020), Green Chem. , 22 : 4450-4455 (2020), etc.) It can be converted to, and these lactone compounds are also described in various previously known literature (Rankic et al. , J. Org. Chem. , 82(23): 12791-12797 (2017), Decker et al. , Tetrahedron, 60 ( 21): 4567-4678 (2004), etc.), it can be converted to a lactam through a known reaction, and can be used in a wider variety of ways.

2. Second embodiment - strategy for mediating asymmetric fatty ether compounds

Another aspect of the present invention provides a method for producing monomers for producing synthetic resins from fatty alcohols.

The method of the present invention includes synthesizing an asymmetric fatty ether compound from fatty alcohol; fermenting the asymmetric fatty ether compound with a genetically recombinant transformant to produce an asymmetric fatty ether compound functionalized at both ends; and decomposing the asymmetric fatty ether compound functionalized at both ends.

1st' stage

First, an asymmetric fatty ether compound is synthesized from fatty alcohol.

The fatty alcohol is a compound in which one terminal of a saturated or unsaturated aliphatic hydrocarbon having m or n carbon atoms is substituted with a hydroxy group. The carbon number m of the hydrocarbon may be an integer of 1 to 20, for example, an integer of 1 to 12, specifically an integer of 1 to 6, especially an integer of 1 to 3. In addition, the carbon number n of the hydrocarbon may be an integer of 3 to 20, for example, an integer of 4 to 18, specifically an integer of 5 to 16, especially an integer of 6 to 14. Additionally, the hydrocarbons may be straight chain or branched, but may especially be straight chain.

The above fatty alcohol can be converted into an asymmetric fatty ether compound through a reaction shown in the following Chemical Formula 5. Through the following process, a fatty alcohol with a carbon number of m and a fatty alcohol with a carbon number of n can be converted into a fat with a carbon number of m + n. Ether compounds may be formed. Here, m and n are different numbers, and the fatty ether compound produced at this time has a hydrocarbon group with m carbon atoms and a hydrocarbon group with n carbon atoms on both sides centered on the O atom of the ether bond, so it is an asymmetric fatty ether compound. It is said that

[Formula 5]

For example, when methanol, an alcohol with an m of 1, reacts with fatty alcohols such as butanol, pentanol, hexanol, heptanol, octanol, nonanol, decanol, undecanol, and dodecanol, each produces methylbutyl ether ( or 1-methoxybutane (C ₅ H ₁₂ O)), methylpentyl ether (1-methoxypentane (C ₆ H ₁₄ O)), methylhexyl ether (1-methoxyhexane (C ₇ H ₁₆ O)). , methylheptyl ether (1-methoxyheptane (C ₈ H ₁₈ O)), methyl octyl ether (1-methoxyoctane (C ₉ H ₂₀ O)), methylnonyl ether (1-methoxynonane (C ₁₀ H ₂₂ O)), methyldecyl ether (1-methoxydecane (C ₁₁ H ₂₄ O)), methyl undecyl ether (1-methoxyundecane (C ₁₂ H ₂₆ O)), methyl dodecyl ether (1- Asymmetric dialkyl ether compounds such as methoxydodecane (C ₁₃ H ₂₈ O)) can be prepared.

In addition, when ethanol, an alcohol with an m of 2, reacts with fatty alcohols such as butanol, pentanol, hexanol, heptanol, octanol, nonanol, decanol, undecanol, dodecanol, etc., ethyl butyl ether ( 1-ethoxybutane (C ₆ H ₁₄ O)), ethylpentyl ether (1-ethoxypentane (C ₇ H ₁₆ O)), ethylhexyl ether (1-ethoxyhexane (C ₈ H ₁₈ O)), Ethylheptyl ether (1-ethoxyheptane (C ₉ H ₂₀ O)), ethyl octyl ether (1-ethoxyheptane (C ₁₀ H ₂₂ O)), ethyl nonyl ether (1-ethoxynonane (C ₁₁ H ₂₄ O)), ethyldecyl ether (1-ethoxydecane (C ₁₂ H ₂₆ O)), ethyl undecyl ether (1-ethoxyundecane (C ₁₃ H ₂₈ O)), ethyl dodecyl ether (1- Asymmetric dialkyl ether compounds such as toxidodecane (C ₁₄ H ₃₀ O)) can be prepared.

Furthermore, even in the case of alcohols with an m of 3 or more, they react with fatty alcohols such as butanol, pentanol, hexanol, heptanol, octanol, nonanol, decanol, undecanol, dodecanol, etc. to form various asymmetric dialkyl ethers. Compounds can be formed.

In addition, Chemical Formula 5 as described above can be performed through methods well known in the technical field to which the present invention pertains, for example, nucleophilic substitution at a temperature of 130 to 140 degrees Celsius using an acid catalyst such as sulfuric acid (H ₂ SO ₄ ). It can be performed by proceeding with the reaction, using dimethyl sulfuric acid ((CH ₃ ) ₂ SO ₄ ) as shown in Formula 6 below, or diethyl sulfuric acid ((CH ₃ CH ₂ ) ₂ SO ₄ ) as shown in Formula 7. It can also be manufactured by reacting with fatty alcohol, but is not limited to this.

[Formula 6]

[Formula 7]

On the other hand, as shown in Table 1 below, when the number of carbon atoms n of the fatty alcohol used for the synthesis of the symmetrical dialkyl ether is 14 or more, the melting point of the produced dialkyl ether compound with 2n carbon atoms exceeds 45 degrees Celsius, so in the present invention, There is a problem in that it is difficult to functionalize both ends through the biological conversion reaction used. As a result, there is a problem in that it is difficult to produce hydroxyalkanoic acids, aminoalkanoic acids, alkanediols, aminoalkanols, diaminoalkanes, etc. with 14 or more carbon atoms only through the strategy of using symmetrical dialkyl ether compounds. However, as shown in Table 1 below, in the case of an asymmetric fatty ether compound, even in the case of ethylhexadecyl ether with a total carbon number of 18 produced using ethanol with m of 2 and hexadecanol with n of 16, the melting point is Celsius. Since the temperature is only about 18 degrees, it is easy for the biological conversion reaction used in the present invention to occur. Therefore, according to the strategy using asymmetrical fatty ether compounds, hydroxyalkanoic acids, aminoalkanoic acids, alkanediols, aminoalkanols, diaminoalkanes, etc. with more carbon atoms can be produced than the strategy using symmetrical dialkyl ether compounds. There is an advantage.

Symmetric dialkyl ether	melting point (temperature in degrees Celsius)	asymmetric fatty ether	melting point (temperature in degrees Celsius)
Dioctyl ether (CH 3 (CH 2 ) 7 -O-(CH 2 ) 7 CH 3 )	-7.6	Ethyl octyl ether (1-Ethoxyoctane) (CH 3 (CH 2 )-O-(CH 2 ) 7 CH 3 )	12.5
Didecyl ether (CH 3 (CH 2 ) 9 -O-(CH 2 ) 9 CH 3 )	16	Ethyldecyl ether (1-Ethoxydecane) (CH 3 (CH 2 )-O-(CH 2 ) 9 CH 3 )	12.5<, <16
Diundecyl ether (CH 3 (CH 2 ) 10 -O-(CH 2 ) 10 CH 3 )	24	Ethyl undecyl ether (1-Ethoxyundecane) (CH 3 (CH 2 )-O-(CH 2 ) 10 CH 3 )
Didodecyl ether (CH 3 (CH 2 ) 11 -O-(CH 2 ) 11 CH 3 )	32	Ethyldodecyl ether (1-Ethoxydodecane) (CH 3 (CH 2 )-O-(CH 2 ) 11 CH 3 )
Ditetradecyl ether (CH 3 (CH 2 ) 13 -O-(CH 2 ) 13 CH 3 )	45	Ethyl tetradecyl ether (1-Ethoxytetradecane) (CH 3 (CH 2 )-O-(CH 2 ) 13 CH 3 )	16
Dihexadecyl ether (CH 3 (CH 2 ) 15 -O-(CH 2 ) 15 CH 3 )	55	Ethylhexadecyl ether (1-Ethoxyhexadecane) (CH 3 (CH 2 )-O-(CH 2 ) 15 CH 3 )	18

2nd' stage

The asymmetric fatty ether compound produced in the first' step is fermented with a genetically recombinant transformant to produce an asymmetric fatty ether compound functionalized at both ends.

This can be performed in the same way as in step 2 of the first embodiment, and once the first genetically recombined transformant is used, both ends are oxidized to oxidize the α, ψ-position of the asymmetric fatty ether compound. A carboxyl group can be introduced.

In addition, using a genetically recombinant second transformant or an appropriate catalyst, the asymmetric fatty ether compound produced as described above whose both ends are functionalized with carboxyl groups can be functionalized at both ends with hydroxy groups.

Furthermore, using a genetically recombinant third transformant or an appropriate catalyst, the asymmetric fatty ether compound produced as described above whose both ends are functionalized with hydroxy groups can be functionalized at both ends with amine groups.

Since the second' step of functionalizing both ends of the asymmetric fatty ether compound is the same biological or chemical method as step 2 of the first embodiment, a detailed description thereof is provided in step 2 of the first embodiment. Parts are used and detailed explanations are omitted.

3rd' stage

The asymmetric fatty ether compound functionalized at both ends produced in the second' step is decomposed. The decomposition also hydrolyzes the ether bond of the asymmetric fatty ether compound functionalized at both ends, and the hydrolysis of the ether bond as described above can be performed through a method well known in the art to which the present invention pertains. , can be performed in the same way as in step 3 of the first embodiment.

Therefore, the hydrolysis of the ether bond as described above can be performed on all types of asymmetric fatty ether compounds functionalized at both ends that can be produced in step 2'. Therefore, by hydrolyzing an asymmetric aliphatic ether compound with m + n carbon atoms functionalized with carboxyl groups at both ends, 1 equivalent of hydroxyalkanoic acid with m carbon atoms and 1 equivalent of hydroxyalkanoic acid with n carbon atoms or amino acid with m carbon atoms are obtained. Produce 1 equivalent of alkanoic acid and 1 equivalent of aminoalkanoic acid with n carbon atoms, or hydrolyze an asymmetric aliphatic ether compound with m+n carbon atoms functionalized with hydroxy groups at both ends to produce 1 equivalent of alkanediol with m carbon atoms and 1 equivalent of carbon alkanoic acid. Generates 1 equivalent of alkanediol with n or 1 equivalent of aminoalkanol with m and 1 equivalent of aminoalkanol with n, or an asymmetric aliphatic ether compound with m+n carbon atoms functionalized with amine groups at both ends. can be hydrolyzed to produce 1 equivalent of an aminoalkanol with m carbon number and 1 equivalent of aminoalkanol with n carbon number, or 1 equivalent of diaminoalkane with m carbon number and 1 equivalent of diaminoalkane with n carbon number.

Various types of monomers such as hydroxyalkanoic acid, aminoalkanoic acid, alkanediol, aminoalkanol, diaminoalkane, etc. prepared as described above can be usefully used to produce synthetic resins such as polyamide and polyester. In particular, hydroxyalkanoic acids are lactone compounds through reactions known in previously known literature (e.g., Pyo et al . (2020), Green Chem. , 22 : 4450-4455 (2020), etc.) It can be converted to, and these lactone compounds are also described in various previously known literature (Rankic et al. , J. Org. Chem. , 82(23): 12791-12797 (2017), Decker et al. , Tetrahedron, 60 ( 21): 4567-4678 (2004), etc.), it can be converted to a lactam through a known reaction, and can be used in a wider variety of ways.

Hereinafter, the present invention will be described in detail through examples.

However, the following examples specifically illustrate the present invention, and the content of the present invention is not limited by the following examples.

[Example 1]

Synthesis of dialkyl ether compounds from fatty alcohols

[1-1] Synthesis of dioctyl ether

The synthesis was carried out by adding sulfuric acid to 0.01 mol% of bio-derived 1-octanol and reacting at 200°C for 20 hours. Dioctyl ether was separated from the reactant through fractional distillation under reduced pressure conditions of 20 torr. The purity of the isolated dioctyl ether was confirmed by a gas chromatography-mass spectrometry system (GC-MS) equipped with a quadrupole electron selective ionization detector (EI) operated at 70 eV. An Agilent HP-5MS column (30 m length, 0.25 mm inner diameter, and 0.25 μm film thickness) was used at a 10:1 split ratio. Helium (flow rate 1.2 mL/min) was used as a carrier gas, and the oven temperature ranged from 100°C to 320°C (10°C/min). The synthesized substrate and an equal volume of diethyl ether were used to extract the substrate, and tetradecane was used as an internal standard. The purity of the synthesized substrate was more than 97% as shown in Figure 2a, and the impurity was unreacted 1-octanol that did not participate in the reaction.

Cell growth toxicity of dioctyl ether synthesized as above was analyzed. Mixed solutions of YPD medium (10g/L yeast extract, 20g/L Bacto peptone, 20g/L glucose) and dioctyl ether at different concentrations (0, 0.25, 0.5, 1, 2, 3, 4, 5) in a 48-well plate. %), add 200㎕ each. Add 2㎕ of culture medium of the recombinant Candida tropicalis Ct6 strain cultured overnight in YPD medium at 30℃ and 200rpm to each well, then culture at 30℃ at 15-minute intervals for more than 24 hours using a plate reader. Measure absorbance at 600 nm. Using the Gompertz growth equation, the maximum specific growth rate ( μ _m ) and lag time were obtained. As a result, as shown in Figure 2b, it was confirmed that the synthesized dioctyl ether had almost no cytotoxicity.

[1-2] Synthesis of didodecyl ether

The synthesis was carried out by adding sulfuric acid to 0.01 mol% of bio-derived 1-dodecanol and reacting at 200°C for 20 hours. Didodecyl ether was separated from the reactant through fractional distillation under reduced pressure conditions of 20 torr. The purity of the isolated didodecyl ether was confirmed by a gas chromatography-mass spectrometry system (GC-MS) equipped with a quadrupole electron selective ionization detector (EI) operated at 70 eV. An Agilent HP-5MS column (30 m length, 0.25 mm inner diameter, and 0.25 μm film thickness) was used at a 10:1 split ratio. Helium (flow rate 1.2 mL/min) was used as a carrier gas, and the oven temperature ranged from 100°C to 320°C (10°C/min). The synthesized substrate and an equal volume of diethyl ether were used to extract the substrate, and tetradecane was used as an internal standard. The purity of the synthesized substrate was over 97% as shown in Figure 2c, and the impurity was unreacted 1-dodecanol that did not participate in the reaction.

The cell growth toxicity of didodecyl ether synthesized as above was analyzed. Add 200㎕ of mixed solution of YPD medium and didodecyl ether of each concentration (0, 0.25, 0.5, 1, 2, 3, 4, 5%) to a 48-well plate. After adding an additional 2㎕ of culture medium of the recombinant Candida tropicalis Ct6 strain cultured overnight in YPD medium at 30℃ and 200rpm to each well, absorbance was measured at 600nm at 33℃ at 15-minute intervals for over 24 hours using a plate reader. Measure. Using the Gompertz growth equation, the maximum growth rate and delay time were obtained. As a result, as shown in Figure 2d, it was confirmed that the synthesized didodecyl ether had almost no cytotoxicity.

[Example 2]

Production of dialkyl ether compounds functionalized with carboxyl groups at both ends from alkyl ether compounds

[2-1] Preparation of α,ψ-carboxylated recombinant strain

Jeon et al . (2019) described in the literature (Jeon et al. , Green Chem , 21, 6491-6501 (2019)), recombinant Candida tropicalis Ct6 strain having the genotype shown in Table 2 below was used. .

recombinant strain	genotype
C. tropicalis Ct6	ura3/ura3 pox2Δ::CYP52A19/POX2 pox4/pox4 pox5Δ::CPRB-URA3/pox5

The Candida tropicalis Ct6 strain was maintained in 15% glycerol at -70 degrees Celsius, and the stock strain was grown on YPD agar medium (10g/L yeast extract, 20g/L Bacto peptone, 20g/L glucose, and 20g/L agarose). The plate was plated and cultured overnight at 30 degrees Celsius.

[2-2] Confirmation of toxicity of α,ψ-carboxylated recombinant strain

Prior to bioconversion of alkyl ether compounds, toxicity tests were performed on α,ψ-carboxylated recombinant strains depending on the concentration of the substrate.

First, YPD medium (10g/L Yeast extract, 20g/L Bacto peptone, 20g/L glucose) and ether compounds with different carbon numbers (dibutyl ether, dipentyl ether, dihexyl ether, diheptyl ether, dioctyl ether, Didecyl ether, diundecyl ether, and didodecyl ether; obtained from each TCI company) were prepared by mixing them at different concentrations (0, 0.25, 0.5, 1, 2, 3, 4, 5%). Then, 200 uL of the mixed solution of YPD and reagent was added to each well of the 48-well plate. Into each well containing the mixed solution, 2 uL of the α,ψ-carboxylated recombinant strain of Example [2-1], which was inoculated in 2 mL YPD medium the day before and cultured at 30 degrees Celsius and 200 rpm for one day, The growth of the α,ψ-carboxylated recombinant strain was measured at 15-minute intervals for 24 hours using a plate reader (Tecan). Using the measured results, a value close to the Gompertz growth equation was obtained, and the maximum specific growth rate (um) and lag time (μ, lag time) were obtained.

As a result, as shown in Figure 3, it was confirmed that ether compounds with 14 or more carbon atoms inhibit the growth of strains less than ether compounds with 12 or less carbon atoms, and in particular, ether compounds with 16 or more carbon atoms decreased by 5%. It was confirmed that cell growth was hardly inhibited even at high substrate concentrations.

[2-3] Confirmation of α,ψ-carboxylation of dialkyl ether compounds

First, to confirm the biological α,ψ-carboxylation of dialkyl ether compounds, a laboratory-scale 5 L culture experiment was conducted. The α,ψ-carboxylated recombinant strain prepared in Example [2-1] was inoculated into 2 mL YPD medium and pre-cultured at 30 degrees Celsius and 200 rpm for 24 hours. The culture medium pre-cultured as above was placed in a 2 L baffled flask containing 200 mL of YPD medium, and cultured at 30 degrees Celsius and 200 rpm for 24 hours. Then, all 200 mL of culture medium cultured the previous day was added to a fermenter containing 1.8 L of medium containing the same ingredients as shown in Table 3 below to make the final volume 2 L. At this time, the final composition of the 2 L culture medium is shown in Table 3 below.

ingredient	Concentration (g/L)
Glycerol	80
CaCl 2 ·2H 2 O	0.1
NaCl	0.1
Yeast extract	20
(NH 4 ) 2 SO 4	8
KH 2 PO 4	2
Trace element	1mL
Antifoam	0.3mL
MgSO 4 ·7H 2 O	One

When all of the glycerol contained in the initial culture was consumed, the feeding medium (600 g/L glucose, 30 g/L NH ₄ Cl) was continuously added at a rate of 0.167 mL/min. To activate the ψ-oxidation pathway of the α,ψ-carboxylated recombinant strain, 3 mL of dodecane (TCI) was added along with the feeding medium. 3 hours after dodecane supply, 10 mL of substrate (dibutyl ether, dipentyl ether, dihexyl ether, diheptyl ether, dioctyl ether, didecyl ether, diundecyl ether, didodecyl ether) was added to the culture medium for reaction. proceeded. The pH was maintained at 6 for initial cell growth, and when the substrate was added, the pH was adjusted to 7.5. The pH of the culture medium was adjusted using 6N NaOH solution. Dissolved oxygen was adjusted by adjusting the stirring speed and maintained above 30%, and the temperature of the fermenter was maintained at 30 degrees Celsius (33 degrees Celsius for diundecyl ether and didodecyl ether). 4 After adding the substrate The culture medium was treated with acid after 6 hours and an equal volume of diethyl ether was used to extract the substrate and product for GC-MS analysis. The substrate and product were then analyzed using a gas chromatography-mass spectrometry (GC-MS) system equipped with a quadrupole electron selective ionization detector operated at 70 eV, using an Agilent HP-5MS column (30 m long, i.d. 0.25 mm and a film thickness of 0.25 um) was used at a split ratio of 10:1. Helium (flow rate 1.2 mL/min) was used as a carrier gas, and the oven temperature was 100 to 320 degrees Celsius (10 degrees Celsius/min). Tetradecane was used as an internal standard, and the carboxylated dialkyl ether compound was inferred from the fragmentation profile of GC-MS because there was no commercially available reagent.

As a result, it was confirmed that all dialkyl ether compounds used as substrates were converted into α,ψ-carboxylation products corresponding to each compound, and the conversion rates were shown in Table 4 below.

temperament	Conversion rate (%)
	After 4 hours of hardening after adding the substrate	6 hours after substrate addition
Dibutyl ether (C8)	65.91 ± 0.17	67.66 ± 0.52
Dipentyl ether (C10)	44.80 ± 0.31	70.79 ± 0.23
Dihexyl ether (C12)	54.54 ± 1.06	73.14 ± 0.55
Diheptyl ether (C14)	80.66 ± 4.89	100
Dioctyl ether (C16)	90.17 ± 0.62	100
Didecyl ether (C20)	65.37 ± 1.35	76.24 ± 1.42
Diundecyl ether (C22)	97.31 ± 1.44	100
Didodecyl ether (C24)	75.58 ± 3.00	100

[2-4] Confirmation of α,ψ-carboxylation of bio-derived dialkyl ether compounds Next, bio-derived dialkyl ether compounds were also subjected to α,ψ-carboxylation recombination prepared in Example [2-1]. In order to confirm whether α,ψ-carboxylation can be biologically performed by the strain, using the dioctyl ether and didodecyl ether synthesized in Example 1 as substrates, Example [2-3] and α,ψ-carboxylation of dialkyl ether compounds was performed in the same manner.

As a result, 160 g/L of α,ψ-carboxylated dioctyl ether after 120 hours as shown in Figure 4a, and 110 g/L of α,ψ after 85 hours as shown in Figure 4b. -Carboxylated didodecyl ether was confirmed to be produced, respectively.

[Example 3]

Production of dialkyl ether compounds functionalized at both ends with hydroxy groups from dialkyl ether compounds functionalized at both ends with carboxyl groups

[3-1] Preparation of α,ψ-hydroxylated recombinant strain

The CAR (carboxylic acid reductase) gene (WP_00582584.1) from Mycobacterium abscessu and the Sfp (4'-phosphopantetheinyl transferase) gene (WP_00234549.1) from Bacillus sp . In order to locate the operon structure under the promoter, the base sequence of SEQ ID No. 1 was requested to be synthesized by ATUM, an American company, and this synthesized gene was inserted into the BamHI and HindIII sites of pJ281 (ATUM). The recombinant vector pJ281-CAR-Sfp as described above was transfected into Escherichia coli MG1665(DE3) strain (Novagen) to produce an α,ψ-hydroxylated recombinant strain.

The α,ψ-hydroxylated recombinant E. coli strain was maintained in 15% glycerol at -70 degrees Celsius, and the stock strain was grown on LB agar medium containing kanamycin (5g/L yeast extract, 10g/L Bacto tryptone, 10g/L L NaCl, 20 g/L agarose, and 50 mg/L Kanamycin) and cultured overnight at 37 degrees Celsius.

[3-2] Confirmation of α,ψ-hydroxylation of dialkyl ether compounds functionalized with carboxyl groups at both ends

Next, in order to confirm the biological α,ψ-hydroxylation of dialkyl ether compounds functionalized with carboxyl groups at both ends, the dioctyl ether synthesized in Example 1 in Examples [2-4] was bioconverted. One culture was centrifuged to remove cells, and then the supernatant was used as a substrate solution to confirm the formation of α,ψ-hydroxylated dioctyl ether.

For this purpose, the α,ψ-hydroxylated recombinant strain prepared in Example [3-1] was cultured in 2 mL LB medium containing kanamycin (5 g/L yeast extract, 10 g/L Bacto tryptone, 10 g/L NaCl, and 50 mg/L L Kanamycin) and pre-cultured for one day at 37 degrees Celsius and 200 rpm. The culture medium pre-cultured as above was placed in a 2L baffled flask containing 200mL LB medium containing kanamycin, and cultured for one day at 37 degrees Celsius and 200 rpm. Then, all 200 mL of the culture medium cultured the previous day was added to the fermenter containing 1.8 L of the medium containing the ingredients shown in Table 5 below to make the final volume 2 L. At this time, the final composition of the 2 L culture medium is shown in Table 5 below.

ingredient	Concentration (g/L)
Glucose	15
Galactose	15
Glycerol	80
Soy peptone	10
Yeast extract	10
(NH 4 ) 2 SO 4	5
KH 2 PO 4	6
Trace element	1mL
Antifoam	0.3mL
MgSO 4 ·7H 2 O	2
Kanamycin	0.05

When all carbon sources such as glucose and glycerol contained in the initial culture medium are consumed, the supply medium (800 g/L glucose, 30 g/L NH ₄ Cl) and the substrate (α,ψ-carboxyl produced in Example [2-4] Sylated dioctyl ether) was continuously added at a rate of 0.167 mL/min. For initial cell growth, the pH was maintained at 7 using ammonia water, and when the substrate was added, the pH was maintained at 7.5 using 6N NaOH. Dissolved oxygen was adjusted by adjusting the stirring speed and maintained above 30%, and the temperature of the fermenter was maintained at 35 degrees Celsius, and the components of the culture solution were analyzed in the same manner as in Example [2-3]. As a result, As shown in Figure 5, it was confirmed that both ends of α,ψ-carboxylated dioctyl ether were hydroxylated to produce α,ψ-hydroxylated dioctyl ether.

[Example 4]

Production of dialkyl ether compounds functionalized at both ends with amine groups from dialkyl ether compounds functionalized at both ends with hydroxy groups

[4-1] Preparation of α,ψ-aminated recombinant strain

ADH (alcohole dehydrogenase) gene (WP_130157107.1) from Aeribacillus pallidus , ψ-TA (ψ-transaminase) gene (WP_011135573.1) from Chromobacterium violaceum, and The AlaDH (alanine dehydrogenase) gene (WP_003243280.1) derived from Bacillus subtilis was requested to be synthesized in an operon structure in the form of the base sequence of SEQ ID NO. ) was inserted into the SmaI and HindIII sites. The recombinant vector as described above was transfected into Escherichia coli MG1665(DE3) strain (Novagen) to produce an α,ψ-aminated recombinant strain.

The α,ψ-aminated recombinant E. coli strain was maintained in 15% glycerol at -70 degrees Celsius, and the stock strain was grown on LB agar medium containing kanamycin (5g/L yeast extract, 10g/L Bacto tryptone, 10g/L NaCl, 20 g/L agarose, and 50 mg/L Kanamycin) and cultured overnight at 37 degrees Celsius.

[4-2] Confirmation of α,ψ-amination of dialkyl ether compounds functionalized with hydroxy groups at both ends

Next, in order to confirm the validity of biological α,ψ-amination of dialkyl ether compounds functionalized with hydroxy groups at both ends, α,ψ-hydroxylated dioctyl was bioconverted in Example [3-2]. The culture solution in which ether was produced was centrifuged to remove the supernatant, and then methanol and water were sequentially added to the precipitate to induce crystallization of α,ψ-hydroxylated dioctyl ether. Then, the crystals filtered through cellulose paper were dissolved in methanol again and 0.5% activated carbon was added to induce decolorization. Then, water was added to induce recrystallization, then filtered again with cellulose paper, washed with distilled water, and dried at 80 degrees Celsius to separate and purify the α,ψ-hydroxylated dioctyl ether to a purity of over 99%. did. Then, using this as a substrate, it was confirmed whether α,ψ-aminated dioctyl ether was produced.

For this purpose, the α,ψ-aminated recombinant strain prepared in Example [4-1] was batch cultured in a 5L fermenter with a medium containing the ingredients shown in Table 5 below, then the culture was centrifuged and the supernatant was removed. , cells were stored at -70 degrees Celsius. Using the strain stored as above, a bioconversion mixture was prepared by dissolving the composition shown in Table 6 in 0.1M phosphate buffer (pH 8.0), and 50 mL of the bioconversion mixture was heated to 37 degrees Celsius in a 500 mL baffled flask for 24 hours. While culturing for more than an hour, the components of the culture solution were analyzed in the same manner as in Example [2-3].

ingredient	Concentration (g/L)
α,ψ-aminated recombinant strain (DCW)	70
NH 4 Cl	7.5
DMSO	20% ( v / v )
L-Alanine	6.7
Benzylamine	6.7
α,ψ--Hydroxylated dioctyl ether	10

As a result, as shown in Figure 6, it was confirmed that both ends of the α,ψ-hydroxylated dioctyl ether were aminated to produce α,ψ-aminated dioctyl ether.

[Example 5]

Hydrolysis of dialkyl ether compounds functionalized at both ends

10 g of the α,ψ-carboxylated dioctyl ether produced in Example [2-4] and the α,ψ-hydroxylated dioctyl ether produced in Example [3-2] were dissolved in 1N sulfuric acid. After dissolving to /L and reacting at 200 degrees Celsius for more than 1 hour using a high pressure reactor, the components of the reactant were analyzed in the same manner as in Example [2-3].

In the case of dioctyl ether, the culture medium in which the dioctyl ether synthesized in Example 1 was bioconverted was centrifuged to remove cells, and then H ₂ SO ₄ was added to the supernatant to lower the pH to 4 and α,ψ-carboxylated Precipitation of dioctyl ether was induced. Then, it was filtered through cellulose paper to obtain precipitated α,ψ-carboxylated dioctyl ether. Acetic acid was added to the α,ψ-carboxylated dioctyl ether obtained as above, completely dissolved at 95 degrees Celsius, and then lowered to room temperature to induce crystallization of the α,ψ-carboxylated dioctyl ether. . Then, it was again filtered through cellulose paper and washed with water to separate and purify α,ψ-carboxylated dioctyl ether with a purity of 86.79%. And this was used as a substrate for hydrolysis.

As a result, 8-hydroxyoctanoic acid was obtained from α,ψ-carboxylated dioctyl ether as shown in Figure 7, and from α,ψ-hydroxylated dioctyl ether as shown in Figure 8. It was confirmed that 1,8-octanediol was produced respectively.

Hydrolysis of α,ψ-aminated dioctyl ether was carried out in Example 4 by adding sulfuric acid to a concentration of 1N to the bioconversion mixture containing α,ψ-aminated dioctyl ether and then using a high pressure reactor. After reacting at 200 degrees Celsius for more than 1 hour, the components of the reactant were analyzed in the same manner as in Example [2-3]. As a result, it was confirmed that 8-aminooctanol was produced.

[Example 6]

Preparation of monomers for producing synthetic resins from asymmetric dialkyl ether compounds

[6-1] Synthesis of ethyldodecyl ether (1-ethoxydodecane)

Toluene, bio-derived 1-dodecanol, and NaOH were mixed at room temperature, reacted at 80 degrees for 1 hour, and then cooled. Diethyl sulfate ((CH ₃ CH ₂ ) ₂ SO ₄ ) was added at 30 degrees Celsius and mixed for 1 hour, then the temperature was raised to 60 degrees Celsius and reacted for 6 hours. The reaction product was filtered to remove salts and then distilled to obtain ethyldodecyl ether (1-ethoxydodecane).

The purity of the product was confirmed using a gas chromatography-mass spectrometry (GC-MS) system equipped with a quadrupole electron selective ionization detector operated at 70 eV, using an Agilent HP-5MS column (30 m long, 0.25 mm i.d. and Film thickness of 0.25 μm) was used at a 10:1 split ratio. Helium (flow rate 1.2 mL/min) was used as a carrier gas, and the oven temperature was 100 to 320 degrees Celsius (10 degrees Celsius/min). The synthesized substrate and an equal volume of diethyl ether were used to extract the substrate, and tetradecane was used as an internal standard. The purity of the synthesized ethyldodecyl ether was over 97%, and the impurity was unreacted 1-decanol that did not participate in the reaction.

[6-2] Production of an asymmetric dialkyl ether compound with both ends functionalized with carboxyl groups from ethyldodecyl ether

In the same manner as in Example [2-3], while supplying ethyldodecyl ether prepared in Example [5-1] as a substrate, α,ψ-carboxylic acid prepared in Example [2-1] was used as a substrate. Fermentation was performed with a live recombinant strain.

After adding ethyldodecyl ether, a substrate, the culture medium was treated with acid, and an equal volume of diethyl ether was used to extract the substrate and product for GC-MS analysis. The substrate and product were then analyzed using a gas chromatography-mass spectrometry (GC-MS) system equipped with a quadrupole electron selective ionization detector operated at 70 eV, using an Agilent HP-5MS column (30 m long, i.d. 0.25 mm and a film thickness of 0.25 um) was used at a split ratio of 10:1. Helium (flow rate 1.2 mL/min) was used as a carrier gas, and the oven temperature was 100 to 320 degrees Celsius (10 degrees Celsius/min). Tetradecane was used as an internal standard, and the carboxylated dialkyl ether compound was inferred from the fragmentation profile of GC-MS because there was no commercially available reagent.

As a result, as shown in Figure 9, it was confirmed that carboxymethyl 11-carboxyundecyl ether (=12-(carboxymethoxy)dodecanoic acid), a compound in which both terminals of ethyldodecyl ether were carboxylated, was produced. It has been done.

[6-3] Utilization of asymmetric dialkyl ether compounds functionalized with carboxyl groups at both ends

Ethyldodecyl ether (=carboxymethyl 11-carboxyundecyl ether), both terminals of which were functionalized with a carboxyl group, produced in Example [5-2], was prepared at both ends in the same manner as in Examples 3 and 4. It can be further functionalized with a hydroxy group or an amine group, and further, an asymmetric dialkyl ether compound in which both terminals are functionalized with a carboxyl group, a hydroxy group, or an amine group is subjected to hydrolysis in the same manner as in Example 5 above to produce 12- A variety of monomers for plastics, such as hydroxydodecanoic acid, 12-aminododecanol, and 1,12-dodecanediol, can be produced.

Meanwhile, in the case of 12-hydroxydodecanoic acid produced as above, lactone is formed through a reaction known in Pyo et al . (2020) (Pyo et al. , Green Chem. , 22: 4450-4455 (2020)) For example, 12-hydroxydodecanoic acid is reacted with thionyl chloride in a suitable solvent such as dichloromethane or chloroform to convert to the corresponding acid chloride, and the resulting acid chloride solution is mixed with triethylamine, pyridine, etc. After neutralizing the hydrogen chloride produced during the reaction by treatment with a suitable base such as triethyl orthoformate or diisopropyl azodicarboxylate etc., a solution of a suitable lactone occluding agent such as triethyl orthoformate or diisopropyl azodicarboxylate is added to the reaction mixture and the mixture is allowed to sit at room temperature for some time. Stir. Then, the mixture is heated under reflux for a considerable period of time at a temperature of about 80 to 100 degrees Celsius to promote lactonization of the acid chloride to form lactone. After completion of the reaction, the reactant is cooled and the unreacted acid chloride is dissolved in water or an appropriate aqueous solution. Add and quench. High-purity dodecane lactone can be produced by extracting dodecane lactone from the reaction mixture using a suitable solvent such as ethyl acetate or dichloromethane and purifying it by distillation or recrystallization.

Furthermore, dodecane lactone produced as described above has been described in various prior literature (Rankic et al. , J. Org. Chem. , 82(23): 12791-12797 (2017), Decker et al. , Tetrahedron, 60(21) ): 4567-4678 (2004), etc.) can be converted to dodecalactam through a known reaction, for example, dodecane lactone is dissolved in an appropriate solvent such as methanol or ethanol, and ammonium hydroxide solution is added to the dodecane lactone solution. The mixture is then stirred at room temperature for a significant period of time, and then the reaction mixture is heated under reflux conditions at a temperature of about 120 to 140 degrees Celsius for a period of time to promote cyclization of the lactone to form the lactam. When the reaction is completed, the mixture is cooled, the pH is adjusted to about 7 using hydrochloric acid, and then dodecane lactam is extracted from the reaction mixture using a suitable solvent such as ethyl acetate or dichloromethane, and purified by recrystallization or chromatography to obtain high purity. of dodecane lactam can be produced.

Although preferred embodiments of the present invention have been described above by way of example, the scope of the present invention is not limited to the specific embodiments described above, and those skilled in the art will understand that the scope of the present invention is within the scope stated in the claims of the present invention. It will be possible to change it appropriately.

The present invention relates to a method of producing various monomers used in the production of synthetic resins from fatty alcohols.

Claims (9)

1. Fermenting the dialkyl ether compound with a genetically recombinant transformant to produce a dialkyl ether compound functionalized at both ends; and

   Decomposing the dialkyl ether compound functionalized at both ends;

   A method of producing a monomer for producing a synthetic resin, including.
2. Producing an asymmetric fatty ether compound functionalized at both ends by fermenting the asymmetric fatty ether compound with a genetically recombinant transformant; and

   Decomposing the asymmetric fatty ether compound functionalized at both ends;

   A method of producing a monomer for producing a synthetic resin, including.
3. In claim 1 or claim 2,

   A method of producing a monomer for producing a synthetic resin, wherein the dialkyl ether compound or the asymmetric fatty ether compound is synthesized from fatty alcohol.
4. In claim 3,

   A method of producing a monomer for producing a synthetic resin, wherein the fatty alcohol is a straight-chain fatty alcohol having 6 to 20 carbon atoms.
5. In claim 1,

   The dialkyl ether compounds include dihexyl ether (C ₁₂ H ₂₆ O), diheptyl ether (C ₁₄ H ₃₀ O), dioctyl ether (C ₁₆ H ₃₄ O), dinonyl ether (C ₁₈ H ₃₈ O), At least one selected from the group consisting of didecyl ether (C ₂₀ H ₄₂ O), diundecyl ether (C ₂₂ H ₄₆ O), and didodecyl ether (C ₂₄ H ₅₀ O),

   The asymmetric fatty ether compounds include methylbutyl ether (or 1-methoxybutane (C ₅ H ₁₂ O)), methylpentyl ether (1-methoxypentane (C ₆ H ₁₄ O)), and methylhexyl ether (1 -Methoxyhexane (C ₇ H ₁₆ O)), methylheptyl ether (1-methoxyheptane (C ₈ H ₁₈ O)), methyl octyl ether (1-methoxyoctane (C ₉ H ₂₀ O)), methyl Nonyl ether (1-methoxynonane (C ₁₀ H ₂₂ O)), methyldecyl ether (1-methoxydecane (C ₁₁ H ₂₄ O)), methyl undecyl ether (1-methoxyundecane (C ₁₂ H ₂₆ O)), methyldodecyl ether (1-methoxydodecane (C ₁₃ H ₂₈ O)), ethylbutyl ether (1-ethoxybutane (C ₆ H ₁₄ O)), ethylpentyl ether (1- Toxypentane (C ₇ H ₁₆ O)), ethylhexyl ether (1-ethoxyhexane (C ₈ H ₁₈ O)), ethylheptyl ether (1-ethoxyheptane (C ₉ H ₂₀ O)), ethyl octyl ether (1-ethoxyheptane (C ₁₀ H ₂₂ O)), ethylnonyl ether (1-ethoxynonane (C ₁₁ H ₂₄ O)), ethyldecyl ether (1-ethoxydecane (C ₁₂ H ₂₆ O)) , ethyl undecyl ether (1-ethoxyundecane (C ₁₃ H ₂₈ O)), ethyl dodecyl ether (1-ethoxydodecane (C ₁₄ H ₃₀ O)), at least one selected from the group consisting of A method for producing monomers for producing synthetic resins.
6. In claim 1 or claim 2,

   The dialkyl ether compound or asymmetric fatty ether compound functionalized at both ends is for producing a synthetic resin, wherein both ends of the dialkyl ether compound or asymmetric fatty ether compound are substituted with any one of a carboxyl group, a hydroxy group and an amine group. How to prepare monomers.
7. In claim 1 or claim 2,

   The genetically recombinant transformant includes a microorganism in which the β-oxidation pathway is blocked and CYP450, CPRb (CYP450 reductase complex), and FAO (fatty alcohol oxidase) are overexpressed.
8. In claim 1 or claim 2,

   A method of producing a monomer for producing a synthetic resin, wherein the decomposition is hydrolysis.
9. In claim 1 or claim 2,

   A method for producing a monomer for producing a synthetic resin, wherein one end of the aliphatic alkane is substituted with any one of a carboxyl group, a hydroxy group, and an amine group, and the other end is substituted with one of a hydroxy group and an amine group.

Images